Rab11 supports amphetamine-stimulated norepinephrine transporter trafficking.
J Neurosci
; 30(23): 7863-77, 2010 Jun 09.
Article
en En
| MEDLINE
| ID: mdl-20534835
ABSTRACT
The norepinephrine transporter (NET) is a presynaptic plasma membrane protein that mediates reuptake of synaptically released norepinephrine. NET is also a major target for medications used for the treatment of depression, attention deficit/hyperactivity disorder, narcolepsy, and obesity. NET is regulated by numerous mechanisms, including catalytic activation and membrane trafficking. Amphetamine (AMPH), a psychostimulant and NET substrate, has also been shown to induce NET trafficking. However, neither the molecular basis nor the nature of the relevant membrane compartments of AMPH-modulated NET trafficking has been defined. Indeed, direct visualization of drug-modulated NET trafficking in neurons has yet to be demonstrated. In this study, we used a recently developed NET antibody and the presence of large presynaptic boutons in sympathetic neurons to examine basal and AMPH-modulated NET trafficking. Specifically, we establish a role for Rab11 in AMPH-induced NET trafficking. First, we found that, in cortical slices, AMPH induces a reduction in surface NET. Next, we observed AMPH-induced accumulation and colocalization of NET with Rab11a and Rab4 in presynaptic boutons of cultured neurons. Using tagged proteins, we demonstrated that NET and a truncated Rab11 effector (FIP2DeltaC2) do not redistribute in synchrony, whereas NET and wild-type Rab11a do. Analysis of various Rab11a/b mutants further demonstrates that Rab11 regulates NET trafficking. Expression of the truncated Rab11a effector (FIP2DeltaC2) attenuates endogenous Rab11 function and prevented AMPH-induced NET internalization as does GDP-locked Rab4 S22N. Our data demonstrate that AMPH leads to an increase of NET in endosomes of single boutons and varicosities in a Rab11-dependent manner.
Texto completo:
1
Bases de datos:
MEDLINE
Asunto principal:
Vesículas Sinápticas
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Proteínas Portadoras
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Adrenérgicos
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Proteínas de Unión al GTP rab
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Proteínas de Transporte de Noradrenalina a través de la Membrana Plasmática
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Anfetamina
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Proteínas de la Membrana
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Neuronas
Límite:
Animals
Idioma:
En
Revista:
J Neurosci
Año:
2010
Tipo del documento:
Article
País de afiliación:
Estados Unidos